Shigeru Itoh

Effects of chloride depletion on electron donation from the water-oxidizing complex to the photosystem II reaction center as measured by the microsecond rise of chlorophyll fluorescence in isolated pea chloroplasts

Abstract The role of Cl − in the electron transfer reactions of the oxidizing side of Photosystem II (PS II) has been studied by measuring the fluorescence yield changes corresponding to the reduction of P + -680, the PS II reaction center chlorophyll, by the secondary PS II donor, Z. In Cl − -depleted chloroplasts, a rapid rise in fluorescence yield was observed following the first and second flashes, but not during the third or subsequent flashes. These results indicate that there exists an additional endogenous electron donor beyond P-680 and Z in Cl − -depleted systems. In contrast, the terminal endogenous donor on the oxidizing side of PS II in Tris-washed preparations has previously been shown to be Z, the component giving rise to EPR signals II f and II vf . The rate of reduction of P + -680 in the Cl − -depleted chloroplasts was as rapid as that measured in uninhibited systems, within the time resolution of our instrument. Again, this is in contrast to Tris-washed preparations in which a dramatic decrease in the rate if this reaction has been previously reported. We have also carried out a preliminary study on the rate of rereduction of Z + in the Cl − -depleted system. Under steady-state conditions, the reduction half-time of Z + in uninhibited systems was about 450 μs, while in the Cl − -depleted chloroplasts, the reduction of Z + was biphasic, one phase with a half-time of about 120 ms, and a slower phase with a half-time of several seconds. The appearance of the quenching state due to P + -680 observed following the third flash on excitation of Cl − -depleted chloroplasts was delayed by two flashed when low concentrations of NH 2 OH (20–50 μM) were included in the medium. Hydrazine at somewhat higher concentrations showed the same effect. This is taken to indicate that the reactions leading to PS II oxidation of NH 2 OH or NH 2 NH 2 are uninhibited by Cl − depletion. Addition of NH 2 OH at low concentrations to Tris-washed chloroplasts did not alter the pattern of the fluorescence yield, indicating that the reactions leading to the NH 2 OH oxidation present in Cl − -depleted systems are absent following Tris inhibition. The results are discussed in terms of an inhibition by Cl − depletion of the reactions of the oxygen-evolving complex. It is suggested that no intermediary redox couple exists between the oxygen-evolving complex and Z, and that Z + is reduced directly by Mn of the complex. In terms of the S-state model, Cl − depletion appears to inhibit the advancement of the mechanism beyond S 2 , but not to inhibit the transitions from S 0 to S 1 , or from S 1 to S 2 .